Amyloid deposition occurs in at least twenty five different human diseases. This project is focused on amyloid formation by Islet Amyloid Polypeptide (IAPP, Amylin), the hormone responsible for pancreatic islet amyloid in type 2 diabetes. IAPP is normally secreted as a soluble polypeptide together with insulin, but aggregates via an unknown mechanism in type 2 diabetes to form extracellular islet amyloid. Islet amyloid, or the process of its formation, is toxic to ?-cells and contributes to the patholog of type 2 diabetes. Amyloid formation by IAPP has been recently shown to be a major complicating factor in islet cell transplantation. Diabetes has reached epidemic proportions in the United States and is emerging as a major health threat in the developing world. Relatively little is known about the mechanism of amyloid formation by IAPP or about the nature of the toxic species generated during its formation. The research will (I) determine the mechanism of amyloid formation by IAPP; (II) examine the effects of a mutation in IAPP which has been proposed to be linked to early onset diabetes; (III) define the most toxic species populated during amyloid assembly and define their conformational properties; (IV) critically test the hypotheses that accelerating amyloid formation reduces IAPP induced cytotoxicity. The research will provide insight into strategies for the treatment of type 2 diabetes and for the prevention of islet graft failure after transplantation. The lessons learned will also aid in the design of more soluble variants of IAPP to treat type 1 diabetes. An interdisciplinary combination of experimental biophysics, biochemistry and cell biology will be used to address these issues. The methods being developed are expected to be broadly applicable and will aid efforts to better control amyloid formation in other diseases. Five specific aims will be carried out. The first involves studies of the mechanism of amyloid formation by IAPP. The second aim will examine the basis for enhanced amyloid formation by a natural mutant of human IAPP. The third aim will interrogate the conformational properties of toxic intermediates populated during amyloid formation by IAPP using high resolution methods, while the fourth aim will conduct comparative studies of toxic and non-toxic IAPP inhibitor complexes. The final aim will test a new strategy for preventing IAPP toxicity.